Rhinovirus and IL-13 alter DNA methylation in pediatric, bronchial epithelial cells at key asthma loci

Organotypic BEC cultures were generated from 10 children with asthma and 6
healthy children from bronchial brushings obtained while donors were
undergoing elective surgery with IRB approval, parent consent, and donor
assent. BEC donors were characterized at an initial research visit to
confirm diagnosis and medical history and to record spirometry and
fraction of exhaled nitric oxide measurements. After differentiation to an
organotypic, pseudostratified state, BEC cultures were infected with RV-16
at an MOI of 0.5 or stimulated with IL-13 at 10 ng/mL. RNA and DNA were
harvested 10 days after RV16 infection and 7 days after IL-13 treatment.
Collected RNA was analyzed via RNA-sequencing for gene expression, and DNA
methylation was quantified in DNA using the Asthma & Allergy
array. Reads were sequenced and aligned to the human genome (GRCh38,
Ensembl 91) with quality control. Genes were filtered to retain only
protein-coding genes and nearest gene annotation for the methylation
dataset, resulting in a final expression dataset with 3,827 genes. Data
are anonymized per IRB protocol and following all ethical and legal
guidelines.